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S Kadis

Publications and source records attributed to S Kadis.

At least 19 recordsLinked to original sources
PubMed1993

Pulmonary lesions in mice inoculated with Actinobacillus pleuropneumoniae hemolysin and lipopolysaccharide.

Non-Swiss albino (CF1) male mice were used as a model to study the effects of specific toxic components of Actinobacillus pleuropneumoniae on the respiratory system. Mice were divided into five groups of ten each and were inoculated by the intranasal route with whole-cell Actinobacillus pleuropneumoniae serotype 1, cell-free culture supernatant fluid (CFCSF) containing hemolysin protein, purified lipopolysaccharide (LPS), heat-treated CFCSF, or phosphate-buffered saline solution. Pulmonary lesions were evaluated at 6 and 12 hours after inoculation. Mice inoculated with whole cells, CFCSF, and LPS developed severe purulent bronchiolitis and alveolitis. Focal pulmonary necrosis was also observed in these three groups but was most consistent in the CFCSF-inoculated mice. Ultrastructurally, lungs from mice inoculated with whole cells, LPS, and CFCSF were characterized by severe degenerative changes in type I and type II pneumocytes. Alveolar spaces contained cellular debris and fibrin. Endothelial cells were swollen, and selected pulmonary capillaries were occluded with platelets and fibrin. Infiltrating neutrophils were often swollen, vacuolated, and degranulated. Although inoculation with relatively large numbers of A. pleuropneumoniae are required to kill mice, the mouse lung appears quite sensitive to the toxic components produced by these bacteria. The elaboration of these two toxic components by A. pleuropneumoniae may be responsible for the characteristic pulmonary inflammatory and necrotic lesions observed in infected swine.

Actinobacillus Infections↗
PubMed1993

Purification of surface-exposed integral outer membrane proteins of Actinobacillus pleuropneumoniae and their role in opsonophagocytosis.

Previously identified 39-, 50-, and 76-kd integral outer membrane proteins (IOMP) of Actinobacillus pleuropneumoniae, a respiratory tract pathogen, were separated by electroelution of sodium dodecyl sulfate-polyacrylamide gel electrophoresis-obtained fragments and their role in opsonophagocytosis by porcine leukocytes was investigated by flow cytometry of fluorescein-labeled A pleuropneumoniae. Using specific antisera, immunoblot analysis indicated that the 3 proteins were antigenically distinct. Antibodies against each IOMP have an important role as opsonins for phagocytosis by porcine leukocytes. The effect of using a combination of all 3 of the specific antisera was minimal. Antiserum absorbed against intact A pleuropneumoniae and Escherichia coli organisms indicated that the antibodies to the 39-, 50-, and 76-kd IOMP were specific for A pleuropneumoniae antigens. Nonheat-treated antiserum did not increase phagocytosis, compared with heat-inactivated antiserum, indicating that complement may not have a major role in opsonization of A pleuropneumoniae.

Actinobacillus pleuropneumoniae↗
PubMed1992

Characterization of lipopolysaccharides from four Pasteurella haemolytica serotype strains: evidence for presence of sialic acid in serotypes 1 and 5.

Highly purified lipopolysaccharides (LPS) obtained from four strains of Pasteurella haemolytica representative of four different serotypes were studied to ascertain their overall structural elements and sugar and fatty acid compositions. SDS-PAGE analysis revealed that each LPS was of the smooth-type although they differed in migration patterns. Somewhat unusual features of these LPS included the presence of: (a) rhamnose in the core oligosaccharides of serotypes 2 and 3; and (b) sialic acid in the LPS of serotypes 1 and 5. The fatty acids, myristic, hydroxymyristic and palmitic occur in essentially equivalent amounts in each of these LPS. In addition, stearic acid was present in small amounts of serotypes 1 and 5.

Amino Sugars↗
PubMed1992

Protective efficacy of conjugate vaccines against experimental challenge with porcine Actinobacillus pleuropneumoniae.

In an attempt to protect pigs against swine pleuropneumonia induced by Actinobacillus pleuropneumoniae (SPAP) by neutralizing the effects of three virulence factors of A. pleuropneumoniae--the capsular polysaccharide (CP), the lipopolysaccharide (LPS), and the hemolysin protein (HP)--two subunit conjugate vaccines were prepared by covalently coupling the CP to the HP and the LPS to the HP. The CP, LPS, and HP were isolated from A. pleuropneumoniae, strain 4074, serotype 1, and the protective efficacy of the conjugate vaccines in swine experimentally infected with A. pleuropneumoniae was evaluated. Following a booster vaccination, a significant (P < 0.05) IgG antibody response to the CP, LPS, and HP was detected in the vaccinated pigs. The pigs vaccinated with the CP-HP and LPS-HP conjugates exhibited significantly less mortality (P < 0.05) and significantly greater weight gain (P < 0.001) than unvaccinated pigs. Vaccinated pigs exhibited significantly fewer and less extensive gross pulmonary lesions (P < 0.001) when compared with unvaccinated pigs. Thus, on the basis of mortality, weight gains, and pulmonary lesion formation, the two conjugate vaccines used in conjunction with one another provide noticeable protective efficacy against SPAP.

Actinobacillus Infections↗
PubMed1992

Effects of Actinobacillus pleuropneumoniae hemolysin on porcine neutrophil function.

In an attempt to gain insight into the events that take place during Actinobacillus pleuropneumoniae infection, the present study was designed to ascertain the effects of bacterial toxicity on porcine neutrophil functions and viability. Incubation of phagocytes (2 x 10(6)) with opsonized A. pleuropneumoniae 4074 (2 x 10(7) CFU) resulted in phagocytic uptake of less than or equal to 4%. At the same bacterium-to-phagocyte ratio, levels of lactate dehydrogenase activity of 74 and 81% were detected in the extracellular medium after 1.5 and 3 h of incubation, respectively. Furthermore, the ingested bacteria were not killed by the phagocytes. These effects were ascribed to hemolysin produced by the bacteria, because the presence of hemolysin-neutralizing antibody prevented overt cellular damage, significantly increased phagocytic uptake (P less than 0.001), and resulted in an approximately 10-fold decrease in the number of CFU of the ingested bacteria. Cytolytic doses of isolated hemolysin caused dose-related loss of cell viability, diminished bactericidal activity of toxin-treated phagocytes for Escherichia coli, and decreased the ability of the phagocytes to undergo a respiratory burst upon stimulation with phorbol myristic acetate. In contrast, sublytic doses of the hemolysin activated the phagocytes and caused them to respond to phorbol myristic acetate with increased generation of superoxide anion. Because heated (100 degrees C, 5 min) hemolysin preparations did not produce similar effects, we contend that the observed effects were not due to contaminating endotoxin. The data presented herein indicate that A. pleuropneumoniae hemolysin is a potent antiphagocytic virulence factor by virtue of its leukocidal activity. Sublytic doses of the toxin may have important effects on the oxidative metabolism of phagocytic cells.

Actinobacillus pleuropneumoniae↗
PubMed1992

Preparation, characterization, and immunogenicity of conjugate vaccines directed against Actinobacillus pleuropneumoniae virulence determinants.

Conjugate vaccines were prepared in an attempt to protect pigs against swine pleuropneumonia induced by Actinobacillus pleuropneumoniae (SPAP). Two subunit conjugates were prepared by coupling the A. pleuropneumoniae 4074 serotype 1 capsular polysaccharide (CP) to the hemolysin protein (HP) and the lipopolysaccharide (LPS) to the HP. Adipic acid dihydrazide was used as a spacer to facilitate the conjugation in a carbodiimide-mediated reaction. The CP and the LPS were found to be covalently coupled to the HP in the conjugates as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and detergent gel chromatography analyses. Following a booster vaccination, pigs exhibited significantly high (P less than 0.05) immunoglobulin G antibodies against CP, LPS, and HP. The anti-CP and anti-LPS immunoglobulin G antibodies were found to function as opsonins in the phagocytosis of A. pleuropneumoniae by polymorphonuclear leukocytes, whereas antibodies to the HP neutralized the cytotoxic effect of the HP on polymorphonuclear leukocytes. No killing of A. pleuropneumoniae was observed when the effects of the antibodies were tested in the presence of complement. Thus, polysaccharide-protein A. pleuropneumoniae conjugates elicit significant antibody responses against each component of each conjugate, which could be instrumental in protecting swine against SPAP.

Actinobacillus pleuropneumoniae↗
PubMed1992

Inhibition of bactericidal activity of anticapsular antibody by nonspecific antibodies reactive with surface-exposed antigenic determinants on Actinobacillus pleuropneumoniae.

In an attempt to understand the mechanism of serum resistance in Actinobacillus pleuropneumoniae, in the present study we examined various interactions among the bacterial surface constituents, serum antibodies, and complement. Analysis of swine sera revealed the presence of anticapsular antibodies in convalescent-phase sera but not in preimmune sera. Both types of sera contained antibodies which reacted with each of 14 polypeptides present in saline extracts of the bacteria. Absorption of the preimmune sera with intact bacteria depleted antibodies to two of the polypeptides (27 and 32 kDa) and high-molecular-weight (greater than 97.4,000) components which did not stain with Coomassie blue. Data derived from complement consumption and C3-binding experiments indicated that the organism was capable of initiating complement activation and binding C3 during incubation in preimmune and immune sera. Experiments designed to evaluate the bactericidal effectiveness of anticapsular antibody revealed that the purified antibody was bactericidal only when preimmune sera absorbed with intact bacteria were used as a source of complement. The bactericidal effects of anticapsular antibody and absorbed preimmune sera were inhibited in a dose-dependent manner by heat-inactivated preimmune sera and immunoglobulin G derived from the sera. The inhibitory activity of the preimmune sera was neutralized by preincubating the sera with column fractions of the saline extract which contained either the 27- or the 32-kDa polypeptide. These results indicate that serum resistance in A. pleuropneumoniae 4074 could be related to inhibition of the bactericidal action of anticapsular antibody by nonspecific antibodies which recognize surface-exposed epitopes on the polypeptides.

Actinobacillus pleuropneumoniae↗
PubMed1991

Immunogenicity of Actinobacillus pleuropneumoniae outer membrane proteins and enhancement of phagocytosis by antibodies to the proteins.

To determine the opsonic effect of antibodies to Actinobacillus (Haemophilus) pleuropneumoniae outer membrane proteins on phagocytosis by porcine polymorphonuclear leukocytes (PMN), we separated the integral outer membrane proteins (IOMPs) by Triton X-114 extraction. Four major IOMPs with molecular masses of 76, 50, 39, and 29 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These IOMPs were found to be essentially free of endotoxin in the Limulus amebocyte lysate assay. The 76-kDa protein exhibited a more intensely stained electrophoresis band when isolated from iron-restricted cultures, and a new band at 105 kDa was present in the whole-membrane fraction but not in the integral fraction, indicating that the 105-kDa iron-repressible protein is a peripheral membrane protein. The 76-, 50-, and 39-kDa proteins were shown to be surface exposed, since antibodies to these IOMPs could be absorbed out of convalescent-phase sera by whole cells. Percentages of phagocytosis by porcine PMN of A. pleuropneumoniae opsonized with convalescent-phase sera, convalescent-phase sera absorbed with IOMPs, or convalescent-phase sera absorbed with whole cells were 46.75, 21.81, and 7.96%, respectively. These results demonstrate that antibodies to IOMPs of A. pleuropneumoniae serve as important opsonins in phagocytosis by porcine PMN.

Actinobacillus↗
PubMed1990

Identification and localization of surface sialylated glycoconjugates in Actinobacillus pleuropneumoniae by direct enzyme-colloidal gold cytochemistry.

Electron microscopic examination of ultrathin sections of 5 strains of Actinobacillus pleuropneumoniae stained for polysaccharides with ruthenium red revealed considerable variability in the amounts of preserved capsular material among the 5 serotypes studied. The amount of capsule was inversely related to the extent of outer membrane-associated sialylated glycoconjugate as evidenced by the degree of binding by colloidal gold-labelled neuraminidase at the cell surface. Serotypes 1, 3, and 5 possessed a well-developed and continuous capsular layer. In serotypes 2 and 7, the capsule consisted of a broken patchy layer that left much of the underlying outer membrane exposed. Morphometric analyses of the mean frequencies of neuraminidase-conjugated gold particles over the perimeters of the A. pleuropneumoniae cells showed that the lowest mean frequencies were observed in serotypes 1, 3, and 5, whereas the second highest and highest mean frequencies were observed in serotypes 7 and 2, respectively. Evidence suggested a serotypic difference in the amount of capsule present and this correlated inversely with the number of sialylated glycoconjugates, which appear to be localized in the outer membranes of the A. pleuropneumoniae cells.

Actinobacillus↗
PubMed1989

Structures and sugar compositions of lipopolysaccharides isolated from seven Actinobacillus pleuropneumoniae serotypes.

Highly purified lipopolysaccharide (LPS) preparations obtained from seven Actinobacillus pleuropneumoniae strains representative of seven different serotypes were used to determine the structure and monosaccharide composition of the polysaccharide components of each lipopolysaccharide. An indication of the structure of each LPS was obtained by procedures that included sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining and gel chromatographic fractionation of acetic acid-hydrolyzed LPS. The polysaccharide components of the LPSs were analyzed by gas-liquid chromatography. The LPSs of the strains of serotypes 2, 4, and 7 were of the smooth type, and those of the strains of serotypes 3 and 6 were of the rough type; the LPSs of the strains of serotypes 1 and 5 could be considered semirough. Rhamnose was present only in the O polysaccharide of the smooth-type and semirough-type LPSs, whereas galactose was present only in the O polysaccharide of the smooth-type LPS and in the core oligosaccharides of the rough-type and semirough-type LPSs. Glucoheptose and mannoheptose were present in the core oligosaccharides of all the LPSs except for the strain of serotype 3, in which only mannoheptose was detected. N-Acetylglucosamine was detected only in the O polysaccharides of the strains of serotypes 1 and 5.

Actinobacillus↗
PubMed1987

Role of haemophilus pleuropneumoniae lipopolysaccharide endotoxin in the pathogenesis of porcine Haemophilus pleuropneumonia.

Intact Haemophilus pleuropneumoniae cells (strain Shope 1, serotype 1), highly purified lipopolysaccharide (LPS) obtained from this strain of H pleuropneumoniae, as well as from Escherichia coli O111:B4, filter-sterilized H pleuropneumoniae cell-free culture supernatant fluid, and heat-inactivated supernatant fluid were given intranasally to CF1 mice and intratracheally to pigs. Pulmonary lesions induced by H pleuropneumoniae in mice were similar to those induced by H pleuropneumoniae in pigs. Histologically, lungs of mice and pigs killed 1 or 2 days after inoculation with 200 micrograms of highly purified H pleuropneumoniae LPS had lesions similar to one another and were similar to those in mice and pigs given intact H pleuropneumoniae, except that little or no necrosis or hemorrhage was observed. In mice killed 1 or 2 days after inoculation of 200 micrograms of E coli O111:B4 LPS, pulmonary lesions were similar to those in mice given H pleuropneumoniae LPS. Pulmonary lesions in mice given cell-free culture supernatant fluid obtained from a midlog-phase growth culture of H pleuropneumoniae cultivated in a chemically defined medium were severe and consisted of neutrophil infiltration and extensive necrosis. In mice, the heat-inactivated supernatant fluid produced mild lesions that consisted of foci of neutrophil aggregation and no necrosis. Extensive necrosis observed in lesions caused by cell-free culture supernatant fluid could be attributed to the action of a heat-labile component, perhaps by the extracellular heat-labile hemolysin produced by H pleuropneumoniae cultivated in chemically defined medium. A LPS endotoxin and a heat-labile factor may be involved in the pulmonary lesion development in the acute phase of porcine Haemophilus pleuropneumonia.

Animals↗
PubMed1986

Isolation, purification, and partial characterization of a lipopolysaccharide from Haemophilus pleuropneumoniae.

Lipopolysaccharide (LPS) from Haemophilus pleuropneumoniae 1536, serotype 2, was isolated and purified by a procedure designed to be equally satisfactory for both smooth- and rough-type LPS. The LPS yield was 53%. Analysis of the preparations revealed that protein, nucleic acid, and cellular phospholipid contamination was negligible (less than 0.1%). Analysis of the sugar content of the LPS by gas-liquid chromatography and colorimetric analysis revealed the presence of rhamnose, mannose, galactose, glucose, heptose, glucosamine, galactosamine, and 3-deoxy-D-manno-2-octulosonic acid. The heptose and glucose contents appeared to be unusually high. The fatty acids of the LPS consisted of a mixture of C14:0 and C16:0 in a ratio of about 4.5:1 (50% of the total) and 3-hydroxy C14:0. When used as a preparatory dose for the dermal Shwartzman reaction, as little as 10 micrograms of the LPS injected intradermally in rabbits produced reddening and swelling. After intravenous injection of a 100-micrograms LPS provoking dose, necrosis was observed at all intradermal injection sites. Limulus amebocyte lysate gelation was observed with an LPS concentration as low as 0.5 ng/ml. A typical biphasic fever response was noted in rabbits injected with as little as 0.25 ng of LPS per kg of body weight.

Animals↗
PubMed1986

Growth and hemolysin production by Haemophilus pleuropneumoniae cultivated in a chemically defined medium.

A chemically defined medium (CDM) has been developed which supports both growth and hemolysin production by Haemophilus pleuropneumoniae. Although the growth rate in stationary cultures was substantially slower in CDM than in trypticase soy broth plus 0.6% yeast extract (TSBYE) and slightly slower than in heart infusion broth (HIB), extracellular hemolysin activity in CDM was slightly higher than in HIB and 16-fold greater than in TSBYE. Maximum hemolytic activity was produced in CDM in early to mid log phase of growth. Hemolytic activity in sterile, cell-free culture supernatant fluids persisted for over 10 days at 4 degrees C and 3-5 days at 37 degrees C, but was completely destroyed at 56 degrees C after 30 min. Total hemolysin inactivation was also achieved in the presence of trypsin or pronase (10 units/mL), but no decrease in hemolytic activity was noted in the presence of DNase or RNase. Iron had little effect on the hemolytic activity in the early stages of growth. However, in the later stages of growth, iron had a pronounced effect with hemolytic activity decreasing as the iron concentration increased from 1 to 500 microM. None of these iron concentrations had any effect on the hemolytic activity when added directly to prepared cell-free culture supernatant fluids. The extracellular hemolysin produced by H. pleuropneumoniae in CDM appears to be a heat-labile protein the activity of which is influenced by iron at certain phases of growth.

Animals↗
PubMed1985

Appearance of immunoglobulin classes and complement (C3) during Corynebacterium renale-induced experimental pyelonephritis in the rat.

The local appearance of various immunoglobulin (Ig) isotypes in the urinary tract during ascending pyelonephritis was studied in rats experimentally infected with Corynebacterium renale. The indirect fluorescent antibody assay was used to detect IgG, IgM, IgA, IgE, and C3 on C renale present in the urine of the experimental animals. Corynebacterium renale coated with IgM and IgG antibodies was found beginning on the 4th day after induced infection, with IgG being the more abundant isotype. Coating with IgA occurred as early as the 4th day, but was less dense than coating with IgG. The presence of C3 on C renale was concurrent with IgM and IgG coating. A significant quantity of IgE could not be identified on antibody-coated C renale. Thus, IgG is the major component of the humoral immune response in this model of ascending pyelonephritis. The IgM early during infection and IgA later during infection seem not to be a major component of the immune response in this model.

Animals↗
PubMed1984

Siderophore production by Proteus mirabilis.

Studies on the isolation and characterization of Proteus mirabilis siderophores provided no evidence that these bacteria synthesize catechol- or hydroxamate-type siderophores. However, gas chromatograph analysis in conjunction with mass spectroscopy revealed the presence of alpha-hydroxyisovaleric acid, a previously unknown metabolite. Additional substantiating evidence for the presence of alpha-hydroxyisovaleric acid in these bacteria was obtained from experiments involving the use of thin-layer chromatography and an ultraviolet absorption spectrum. This compound was found to be capable of removing iron from the synthetic chelator, ethylene-diamine-di-orthohydroxyphenyl acetic acid, and supplying that iron to the bacteria both in a solid agar medium and in a liquid medium. Proteus mirabilis was found to possess an enzyme capable of catalyzing the reaction by which alpha-hydroxyisovaleric acid is converted to alpha-ketoisovaleric acid, an intermediate in the valine biosynthetic pathway.

Animals↗
PubMed1984

Relationship of iron administration to susceptibility of newborn pigs to enterotoxic colibacillosis.

To determine whether supplemental iron (Fe) administration to newborn pigs reared in concrete pens not only prevents anemia, but renders the pigs more susceptible to Escherichia coli-induced diarrheal disease, pigs were given a large or a small dose of Fe IM or orally before or after challenge exposure with E coli. The controls were challenge-exposed pigs not given Fe and pigs not challenge exposed (Fe-treated and nontreated groups). Although the mortality of the pigs challenge exposed with E coli and administered a large oral dose of Fe shortly after birth was greater than that of the challenge-exposed pigs given no Fe, differences in mortality were not noted between any of the groups tested when the Fe was injected IM. The Fe-treated survivors had severe diarrhea (oral Fe administration) or mild diarrhea (IM Fe administration) for longer periods than did the nontreated survivors. All challenge-exposed pigs treated with a large dose of Fe gained less weight than the nontreated pigs during the diarrheal period and for several days thereafter. Beyond this time period, the weight gain of the Fe-treated pigs was substantially greater than that of their nontreated littermates; the weight gain of the pigs given a small dose of Fe was intermediate. Hemoglobin and hematocrit values of the pigs shortly after birth and weekly thereafter revealed that within 2 weeks, both sets of values from the pigs treated with a large dose of Fe were within acceptable laboratory limits and substantially greater than the values obtained for the nontreated pigs, which were severely anemic.

Anemia, Hypochromic↗
PubMed1982

Transfer of drug resistance and enterotoxin production in porcine Escherichia coli strains and relationship between K88 antigen and raffinose (melitose) fermentation.

To obtain an indication of the frequency of drug resistance together with enterotoxin production in a select group of porcine E coli strains, those enterotoxigenic strains that were resistant to specific antimicrobial agents were mated as donors with nonenterotoxigenic E coli strains that were sensitive to these drugs as recipients. Seven of the 17 strains tested transferred drug resistance by direct mating. These 7 strains also transferred drug resistance following mobilization. The frequency of recipient conversion was increased following mobilization, as compared with the frequency of conversion resulting from direct mating. In 2 of these strains, enterotoxin production was transferred together with drug resistance. Eight strains transferred drug resistance only after mobilization, and in 2 of these, enterotoxin production was transferred simultaneously with resistance to the antimicrobial agents. In 2 of the 17 strains tested, neither transfer of drug resistance nor transfer of enterotoxin production could be achieved by direct mating or after mobilization. Thirty-nine porcine E coli strains were tested for K88 antigen production and raffinose (melitose) fermentation with a view toward determining whether raffinose fermentation could be used as a diagnostic tool for the detection of the K88 antigen. In 11 of the K88-positive strains tested, only 7 fermented raffinose. In the remaining 28 K88-negative strains, 17 fermented raffinose. Consequently no meaningful correlation existed between these 2 properties of porcine enterotoxigenic E coli strains.

Animals↗